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Expression, Purification, And Monitoring Of Conformational Changes Of Hcb2 Tmh67h8 In Different Membrane-Mimetic Lipid Mixtures Using Circular Dichroism And Nmr Techniques, Elvis K. Tiburu, Jianqin Zhuang, Heidimarie N. A. Fleischer, Patrick K. Arthur, Gordon A. Awandare
Expression, Purification, And Monitoring Of Conformational Changes Of Hcb2 Tmh67h8 In Different Membrane-Mimetic Lipid Mixtures Using Circular Dichroism And Nmr Techniques, Elvis K. Tiburu, Jianqin Zhuang, Heidimarie N. A. Fleischer, Patrick K. Arthur, Gordon A. Awandare
Publications and Research
This work was intended to develop self-assembly lipids for incorporating G-protein coupled receptors (GPCRs) in order to improve the success rate for nuclear magnetic resonance spectroscopy (NMR) structural elucidation. We hereby report the expression and purification of uniformly 15N-labeled human cannabinoid receptor-2 domain in insect cell media. The domain was refolded by screening several membrane mimetic environments. Different q ratios of isotropic bicelles were screened for solubilizing transmembrane helix 6, 7 and 8 (TMH67H8). As the concentration of dimyristoylphosphocholine (DMPC) was increased such that the q ratio was between 0.16 and 0.42, there was less crowding in the cross peaks …